Héchard Y, Dérijard B, Letellier F, Cenatiempo Y
Institut de Biologie Moléculaire et d'Ingénierie Génétique, URA CNRS 1172, Université de Poitiers, France.
J Gen Microbiol. 1992 Dec;138(12):2725-31. doi: 10.1099/00221287-138-12-2725.
A Leuconostoc mesenteroides ssp. mesenteroides was isolated from goat's milk on the basis of its ability to inhibit the growth of Listeria monocytogenes. The antimicrobial effect was due to the presence in the culture medium of a compound, named mesentericin Y105, excreted by the Leuconostoc mesenteroides Y105. The compound displayed known features of bacteriocins from lactic acid bacteria. It appeared as a proteinaceous molecule exhibiting a narrow inhibitory spectrum limited to genus Listeria. The apparent relative molecular mass, as indicated by activity detection after SDS-PAGE, was 2.5-3.0 kDa. The bacteriocin was purified to homogeneity by a simple three-step procedure: a crude supernatant obtained from an early-stationary-phase culture in a defined medium was subjected to affinity chromatography on a blue agarose column, followed by ultrafiltration through a 5 kDa cut-off membrane, and finally by reverse-phase HPLC on a C4 column. Microsequencing of the pure bacteriocin and of tryptic fragments showed that mesentericin Y105 is a 36 amino acid polypeptide whose primary structure is close to that of leucocin A-UAL 187, which contains an extra residue at the C-terminus and displays only two differences in the overlapping sequence. However, unlike leucocin A-UAL 187, mesentericin Y105 displayed a bactericidal mode of action.
基于其抑制单核细胞增生李斯特菌生长的能力，从山羊奶中分离出了肠系膜明串珠菌肠系膜亚种。抗菌作用归因于肠系膜明串珠菌Y105分泌到培养基中的一种名为肠膜菌素Y105的化合物。该化合物表现出乳酸菌细菌素的已知特征。它呈现为一种蛋白质分子，具有狭窄的抑制谱，仅限于李斯特菌属。经SDS-PAGE后活性检测显示，其表观相对分子质量为2.5 - 3.0 kDa。通过一个简单的三步程序将该细菌素纯化至同质：从限定培养基中早期稳定期培养物获得的粗上清液先在蓝色琼脂糖柱上进行亲和层析，然后通过截留分子量为5 kDa的膜进行超滤，最后在C4柱上进行反相高效液相色谱。对纯细菌素及其胰蛋白酶片段的微量测序表明，肠膜菌素Y105是一种36个氨基酸的多肽，其一级结构与亮菌素A-UAL 187相近，亮菌素A-UAL 187在C端含有一个额外的残基，且在重叠序列中仅显示两个差异。然而，与亮菌素A-UAL 187不同，肠膜菌素Y105表现出杀菌作用模式。
